Document Detail


Immunofluorescent staining of human metaphase chromosomes with monoclonal antibody to histone H2B.
MedLine Citation:
PMID:  6761099     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A mouse monoclonal IgM antibody against the core histone H2B has been shown, by indirect immunofluorescence, to stain metaphase chromosomes from a variety of cultured cell types. Experiments carried out with human HeLa cells showed that the intensity of staining varied along the length of chromosome arms giving in some cases a rudimentary banded staining pattern. Considerable variation in staining intensity was noted between individual chromosomes and between different metaphase spreads. It was noted that chromosomes having a more swollen appearance stained more intensely than those with a more compact structure, which were often unstained. Preincubation of unfixed metaphase chromosomes in buffered salt solutions virtually eliminated the cell to cell and chromosome to chromosome variation in staining, even when no visible effect on chromosome morphology was caused by such treatment. It is concluded that the determinant recognised by antibody HBC-7 is ubiquitous but is inaccessible in some chromosomes or chromosome regions. Digestion of purified chromatin (primarily interphase) with DNAase 1 or micrococcal nuclease resulted in a several-fold increase in the binding of antibody HBC-7 measured by solid-phase radioimmunoassay. This increase was abolished by subsequent treatment with trypsin, which suggests that the antigenic determinant recognised by antibody HBC-7 lies in the trypsin-sensitive N-terminal region of nucleosomal H2B. As the cationic N-terminal regions of the core histones are involved in DNA binding, it is likely that the accessibility of the determinant recognised by antibody HBC-7 is influenced by the relationship between the core histones and their associated DNA.
Authors:
B M Turner
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Chromosoma     Volume:  87     ISSN:  0009-5915     ISO Abbreviation:  Chromosoma     Publication Date:  1982  
Date Detail:
Created Date:  1983-04-15     Completed Date:  1983-04-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985138R     Medline TA:  Chromosoma     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  345-57     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal
Antigens
Chromosomes, Human / metabolism*
Deoxyribonucleases
Fluorescent Antibody Technique
Hela Cells / metabolism
Histones / immunology,  metabolism*
Humans
Metaphase
Sodium Chloride
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens; 0/Histones; 7647-14-5/Sodium Chloride; EC 3.1.-/Deoxyribonucleases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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